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Nucleic Acids Res. Mar 1, 1996; 24(5): 931–937.
PMCID: PMC145711

Molecular cloning and functional analysis of a human cDNA encoding an Escherichia coli AlkB homolog, a protein involved in DNA alkylation damage repair.

Abstract

The Escherichia coli AlkB protein is involved in protecting cells against mutation and cell death induced specifically by SN2-type alkylating agents such as methyl methanesulfonate (MMS). A human cDNA encoding a polypeptide homologous to E.coli AlkB was discovered by searching a database of expressed sequence tags (ESTs) derived from high throughput cDNA sequencing. The full-length human AlkB homolog (hABH) cDNA clone contains a 924 bp open reading frame encoding a 34 kDa protein which is 52% similar and 23% identical to E.coli AlkB. The hABH gene, which maps to chromosome 14q24, was ubiquitously expressed in 16 human tissues examined. When hABH was expressed in E.coli alkB mutant cells partial rescue of the cells from MMS-induced cell death occurred. Under the conditions used expression of hABH in skin fibroblasts was not regulated by treatment with MMS. Our findings show that the AlkB protein is structurally and functionally conserved from bacteria to human, but its regulation may have diverged during evolution.

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Selected References

These references are in PubMed. This may not be the complete list of references from this article.
  • Kanamaru R, Ishioka C, Konishi Y, Ishikawa A, Shibata H, Wakui A. Damage to DNA strand and the effects of anticancer drugs. Sci Rep Res Inst Tohoku Univ Med. 1989 Dec;36(1-4):29–36. [PubMed]
  • Frankfurt OS. Inhibition of DNA repair and the enhancement of cytotoxicity of alkylating agents. Int J Cancer. 1991 Jul 30;48(6):916–923. [PubMed]
  • Potter PM, Wilkinson MC, Fitton J, Carr FJ, Brennand J, Cooper DP, Margison GP. Characterisation and nucleotide sequence of ogt, the O6-alkylguanine-DNA-alkyltransferase gene of E. coli. Nucleic Acids Res. 1987 Nov 25;15(22):9177–9193. [PMC free article] [PubMed]
  • Rebeck GW, Smith CM, Goad DL, Samson L. Characterization of the major DNA repair methyltransferase activity in unadapted Escherichia coli and identification of a similar activity in Salmonella typhimurium. J Bacteriol. 1989 Sep;171(9):4563–4568. [PMC free article] [PubMed]
  • Riazuddin S, Lindahl T. Properties of 3-methyladenine-DNA glycosylase from Escherichia coli. Biochemistry. 1978 May 30;17(11):2110–2118. [PubMed]
  • Bjelland S, Bjørås M, Seeberg E. Excision of 3-methylguanine from alkylated DNA by 3-methyladenine DNA glycosylase I of Escherichia coli. Nucleic Acids Res. 1993 May 11;21(9):2045–2049. [PMC free article] [PubMed]
  • Samson L, Cairns J. A new pathway for DNA repair in Escherichia coli. Nature. 1977 May 19;267(5608):281–283. [PubMed]
  • Lindahl T, Sedgwick B, Sekiguchi M, Nakabeppu Y. Regulation and expression of the adaptive response to alkylating agents. Annu Rev Biochem. 1988;57:133–157. [PubMed]
  • Shevell DE, Friedman BM, Walker GC. Resistance to alkylation damage in Escherichia coli: role of the Ada protein in induction of the adaptive response. Mutat Res. 1990 Nov-Dec;233(1-2):53–72. [PubMed]
  • Samson L. The suicidal DNA repair methyltransferases of microbes. Mol Microbiol. 1992 Apr;6(7):825–831. [PubMed]
  • Evensen G, Seeberg E. Adaptation to alkylation resistance involves the induction of a DNA glycosylase. Nature. 1982 Apr 22;296(5859):773–775. [PubMed]
  • Karran P, Hjelmgren T, Lindahl T. Induction of a DNA glycosylase for N-methylated purines is part of the adaptive response to alkylating agents. Nature. 1982 Apr 22;296(5859):770–773. [PubMed]
  • Nakabeppu Y, Kondo H, Sekiguchi M. Cloning and characterization of the alkA gene of Escherichia coli that encodes 3-methyladenine DNA glycosylase II. J Biol Chem. 1984 Nov 25;259(22):13723–13729. [PubMed]
  • Volkert MR, Nguyen DC, Beard KC. Escherichia coli gene induction by alkylation treatment. Genetics. 1986 Jan;112(1):11–26. [PMC free article] [PubMed]
  • Xiao W, Derfler B, Chen J, Samson L. Primary sequence and biological functions of a Saccharomyces cerevisiae O6-methylguanine/O4-methylthymine DNA repair methyltransferase gene. EMBO J. 1991 Aug;10(8):2179–2186. [PMC free article] [PubMed]
  • Shiraishi A, Sakumi K, Nakatsu Y, Hayakawa H, Sekiguchi M. Isolation and characterization of cDNA and genomic sequences for mouse O6-methylguanine-DNA methyltransferase. Carcinogenesis. 1992 Feb;13(2):289–296. [PubMed]
  • Hayakawa H, Koike G, Sekiguchi M. Expression and cloning of complementary DNA for a human enzyme that repairs O6-methylguanine in DNA. J Mol Biol. 1990 Jun 20;213(4):739–747. [PubMed]
  • Rydberg B, Spurr N, Karran P. cDNA cloning and chromosomal assignment of the human O6-methylguanine-DNA methyltransferase. cDNA expression in Escherichia coli and gene expression in human cells. J Biol Chem. 1990 Jun 5;265(16):9563–9569. [PubMed]
  • Chen J, Derfler B, Maskati A, Samson L. Cloning a eukaryotic DNA glycosylase repair gene by the suppression of a DNA repair defect in Escherichia coli. Proc Natl Acad Sci U S A. 1989 Oct;86(20):7961–7965. [PMC free article] [PubMed]
  • Chen J, Derfler B, Samson L. Saccharomyces cerevisiae 3-methyladenine DNA glycosylase has homology to the AlkA glycosylase of E. coli and is induced in response to DNA alkylation damage. EMBO J. 1990 Dec;9(13):4569–4575. [PMC free article] [PubMed]
  • Berdal KG, Bjørås M, Bjelland S, Seeberg E. Cloning and expression in Escherichia coli of a gene for an alkylbase DNA glycosylase from Saccharomyces cerevisiae; a homologue to the bacterial alkA gene. EMBO J. 1990 Dec;9(13):4563–4568. [PMC free article] [PubMed]
  • O'Connor TR, Laval F. Isolation and structure of a cDNA expressing a mammalian 3-methyladenine-DNA glycosylase. EMBO J. 1990 Oct;9(10):3337–3342. [PMC free article] [PubMed]
  • Engelward BP, Boosalis MS, Chen BJ, Deng Z, Siciliano MJ, Samson LD. Cloning and characterization of a mouse 3-methyladenine/7-methyl-guanine/3-methylguanine DNA glycosylase cDNA whose gene maps to chromosome 11. Carcinogenesis. 1993 Feb;14(2):175–181. [PubMed]
  • Samson L, Derfler B, Boosalis M, Call K. Cloning and characterization of a 3-methyladenine DNA glycosylase cDNA from human cells whose gene maps to chromosome 16. Proc Natl Acad Sci U S A. 1991 Oct 15;88(20):9127–9131. [PMC free article] [PubMed]
  • O'Connor TR, Laval J. Human cDNA expressing a functional DNA glycosylase excising 3-methyladenine and 7-methylguanine. Biochem Biophys Res Commun. 1991 May 15;176(3):1170–1177. [PubMed]
  • Chakravarti D, Ibeanu GC, Tano K, Mitra S. Cloning and expression in Escherichia coli of a human cDNA encoding the DNA repair protein N-methylpurine-DNA glycosylase. J Biol Chem. 1991 Aug 25;266(24):15710–15715. [PubMed]
  • Santerre A, Britt AB. Cloning of a 3-methyladenine-DNA glycosylase from Arabidopsis thaliana. Proc Natl Acad Sci U S A. 1994 Mar 15;91(6):2240–2244. [PMC free article] [PubMed]
  • Yarosh DB. The role of O6-methylguanine-DNA methyltransferase in cell survival, mutagenesis and carcinogenesis. Mutat Res. 1985 Jan-Mar;145(1-2):1–16. [PubMed]
  • Chen J, Samson L. Induction of S.cerevisiae MAG 3-methyladenine DNA glycosylase transcript levels in response to DNA damage. Nucleic Acids Res. 1991 Dec 11;19(23):6427–6432. [PMC free article] [PubMed]
  • Kataoka H, Yamamoto Y, Sekiguchi M. A new gene (alkB) of Escherichia coli that controls sensitivity to methyl methane sulfonate. J Bacteriol. 1983 Mar;153(3):1301–1307. [PMC free article] [PubMed]
  • Chen BJ, Carroll P, Samson L. The Escherichia coli AlkB protein protects human cells against alkylation-induced toxicity. J Bacteriol. 1994 Oct;176(20):6255–6261. [PMC free article] [PubMed]
  • Volkert MR, Hajec LI. Molecular analysis of the aidD6::Mu d1 (bla lac) fusion mutation of Escherichia coli K12. Mol Gen Genet. 1991 Oct;229(2):319–323. [PubMed]
  • Kataoka H, Sekiguchi M. Molecular cloning and characterization of the alkB gene of Escherichia coli. Mol Gen Genet. 1985;198(2):263–269. [PubMed]
  • Kondo H, Nakabeppu Y, Kataoka H, Kuhara S, Kawabata S, Sekiguchi M. Structure and expression of the alkB gene of Escherichia coli related to the repair of alkylated DNA. J Biol Chem. 1986 Nov 25;261(33):15772–15777. [PubMed]
  • Cleaver JE. It was a very good year for DNA repair. Cell. 1994 Jan 14;76(1):1–4. [PubMed]
  • Fishel R, Lescoe MK, Rao MR, Copeland NG, Jenkins NA, Garber J, Kane M, Kolodner R. The human mutator gene homolog MSH2 and its association with hereditary nonpolyposis colon cancer. Cell. 1993 Dec 3;75(5):1027–1038. [PubMed]
  • Leach FS, Nicolaides NC, Papadopoulos N, Liu B, Jen J, Parsons R, Peltomäki P, Sistonen P, Aaltonen LA, Nyström-Lahti M, et al. Mutations of a mutS homolog in hereditary nonpolyposis colorectal cancer. Cell. 1993 Dec 17;75(6):1215–1225. [PubMed]
  • Bronner CE, Baker SM, Morrison PT, Warren G, Smith LG, Lescoe MK, Kane M, Earabino C, Lipford J, Lindblom A, et al. Mutation in the DNA mismatch repair gene homologue hMLH1 is associated with hereditary non-polyposis colon cancer. Nature. 1994 Mar 17;368(6468):258–261. [PubMed]
  • Papadopoulos N, Nicolaides NC, Wei YF, Ruben SM, Carter KC, Rosen CA, Haseltine WA, Fleischmann RD, Fraser CM, Adams MD, et al. Mutation of a mutL homolog in hereditary colon cancer. Science. 1994 Mar 18;263(5153):1625–1629. [PubMed]
  • Nicolaides NC, Papadopoulos N, Liu B, Wei YF, Carter KC, Ruben SM, Rosen CA, Haseltine WA, Fleischmann RD, Fraser CM, et al. Mutations of two PMS homologues in hereditary nonpolyposis colon cancer. Nature. 1994 Sep 1;371(6492):75–80. [PubMed]
  • Adams MD, Kelley JM, Gocayne JD, Dubnick M, Polymeropoulos MH, Xiao H, Merril CR, Wu A, Olde B, Moreno RF, et al. Complementary DNA sequencing: expressed sequence tags and human genome project. Science. 1991 Jun 21;252(5013):1651–1656. [PubMed]
  • Adams MD, Dubnick M, Kerlavage AR, Moreno R, Kelley JM, Utterback TR, Nagle JW, Fields C, Venter JC. Sequence identification of 2,375 human brain genes. Nature. 1992 Feb 13;355(6361):632–634. [PubMed]
  • Altschul SF, Gish W, Miller W, Myers EW, Lipman DJ. Basic local alignment search tool. J Mol Biol. 1990 Oct 5;215(3):403–410. [PubMed]
  • Johnson CV, McNeil JA, Carter KC, Lawrence JB. A simple, rapid technique for precise mapping of multiple sequences in two colors using a single optical filter set. Genet Anal Tech Appl. 1991 Apr;8(2):75–76. [PubMed]
  • Johnson CV, Singer RH, Lawrence JB. Fluorescent detection of nuclear RNA and DNA: implications for genome organization. Methods Cell Biol. 1991;35:73–99. [PubMed]
  • Kozak M. At least six nucleotides preceding the AUG initiator codon enhance translation in mammalian cells. J Mol Biol. 1987 Aug 20;196(4):947–950. [PubMed]
  • Proudfoot NJ, Brownlee GG. 3' non-coding region sequences in eukaryotic messenger RNA. Nature. 1976 Sep 16;263(5574):211–214. [PubMed]
  • Volkert MR, Nguyen DC. Induction of specific Escherichia coli genes by sublethal treatments with alkylating agents. Proc Natl Acad Sci U S A. 1984 Jul;81(13):4110–4114. [PMC free article] [PubMed]
  • Lefebvre P, Zak P, Laval F. Induction of O6-methylguanine-DNA-methyltransferase and N3-methyladenine-DNA-glycosylase in human cells exposed to DNA-damaging agents. DNA Cell Biol. 1993 Apr;12(3):233–241. [PubMed]
  • Fritz G, Tano K, Mitra S, Kaina B. Inducibility of the DNA repair gene encoding O6-methylguanine-DNA methyltransferase in mammalian cells by DNA-damaging treatments. Mol Cell Biol. 1991 Sep;11(9):4660–4668. [PMC free article] [PubMed]
  • Lawrence JB, Villnave CA, Singer RH. Sensitive, high-resolution chromatin and chromosome mapping in situ: presence and orientation of two closely integrated copies of EBV in a lymphoma line. Cell. 1988 Jan 15;52(1):51–61. [PubMed]
  • Stevanin G, Le Guern E, Ravisé N, Chneiweiss H, Dürr A, Cancel G, Vignal A, Boch AL, Ruberg M, Penet C, et al. A third locus for autosomal dominant cerebellar ataxia type I maps to chromosome 14q24.3-qter: evidence for the existence of a fourth locus. Am J Hum Genet. 1994 Jan;54(1):11–20. [PMC free article] [PubMed]
  • Wei YF, Chen BJ, Samson L. Suppression of Escherichia coli alkB mutants by Saccharomyces cerevisiae genes. J Bacteriol. 1995 Sep;177(17):5009–5015. [PMC free article] [PubMed]
  • Fornace AJ, Jr, Alamo I, Jr, Hollander MC. DNA damage-inducible transcripts in mammalian cells. Proc Natl Acad Sci U S A. 1988 Dec;85(23):8800–8804. [PMC free article] [PubMed]

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